Oral Immunization for Anti-snake Venom Production in Equine Animals

The immunization process of current commercial manufacturing of anti-snake venom (ASV), uses injections of bentonite, complete Freund’s adjuvant, or incomplete Freund’s adjuvant, mixed with low doses of the snake venom in horses (but rarely in other large mammals), which frequently cause serious adverse effects in host animals. At the site of injection, horses may develop painful swelling, granuloma, abscess, scar, or systemic neurological and hematological defects, low antibody response, or death due to anaphylactic shock. We sought to investigate a novel alternate immunization strategy with oral administration of snake venom with adjuvants. We utilized M5904 mineral oil emulsion as an adjuvant that was mixed with sub-lethal doses (LD) of the snake venoms. Our preliminary experiments were initiated in March 2011 and the present data culminated in March 2018. In our initial experiments which were carried out in inbred mice, the LD100 was 10.36 ug/25 grams of mice for Naja. oxins and 10.0 ug/25 gram of Naja. karachians. We extrapolated the sub-LD dose to horses by cutting the LD100 in mice to 20%. This dose did not cause any apparent pathology in horses and therefore, we adopted that dose for the equine.

ratio of smear-negative tuberculosis in Sindh: an insight using fluorescence microscopy

Background: Sputum smear microscopy is the most useful tool used for the screening of presumptive tuberculosis cases. Conventionally Ziehl-Neelsen [ZN] based staining is used to microscopically screen sputum from the presumptive Tuberculosis Cases. However, Fluorescent microscopy [FM] is now gradually replacing the conventional ZN microscopy because of its higher efficiency and efficacy in detecting Mycobacterium tuberculosis. In patients, who do not produce sufficiently high number of bacilli, and are known as Smear-negative, efficacy of both types of microscopy remains questionable as no Mycobacteria are detected in the sputum of these patients and they are falsely labelled as smear-negative

Objective: This retrospective study was conducted on the patient samples received from different parts of Sindh to study the proportions of false-negative tuberculosis using fluorescent microscopy

Materials and Methods: A retrospective cross-sectional study was conducted from June 2014 to June 2016 at the Tuberculosis Laboratories of Dow University of Health Sciences. Data analyzed for 1439 samples received during 24-month period, which had subsequently been confirmed by culture. All laboratory work was performed as per guidelines of the National Program. Statistical analysis was performed on MedCalc[registered sign] and Open Epi[registered sign]

Results: During 24-month period, among the 1439 cases, 832[57.81%] were from males and 499 [34.67%] were smear negative. Culture showed that 72 [14.42%] were false-negative among all-negatives. Overall sensitivity observed was 92.21% [95%CI 90.29%-93.85%] and specificity 82.91% [95%CI 79.38%- 86.06%]. The proportion of false negativity among all negatives in females [16.11%] was significantly higher [p=0.002] as compared to males [13.47%]

Conclusions: Even with the introduction of Fluorescent Microscopy, smear microscopy cannot be solely relied upon for the diagnosis of tuberculosis, as considerable number of patients producing insufficient number of bacilli to be detected by microscopy

In vitro evaluation of antimicrobial effect of miswak against common oral pathogens

Miswak is a natural tooth cleaning tool which is being used in many parts of the world since ancient times. It is known to be useful in prevention of dental caries. But still it is not used as frequently as other oral hygiene tools. This research was designed to scientifically establish antimicrobial effect of miswak in vitro against common oral pathogens. This was a cross-sectional study involving 100 health care workers. This research was carried out in Microbiology section of Dow Diagnostic Research and Reference Laboratory. A questionnaire was designed to test oral hygiene habits of study subjects. Oral swabs were taken and microorganisms were identified by standard bacteriological methods. Test material included four different types of miswaks i.e. [1] root of the peelu [Salvadora persica] tree [in packing] [2] root of the peelu tree [without packing] [3] stem of the peelu tree and [4] stem of the neem [Azadirechta indica] tree. These miswaks were tested against three different types of microorganisms isolated from oral swabs: Staphylococcus aureus, Streptococcus mutans and Candida albicans by agar diffusion method. Inhibition zone was measured after 24 hrs of incubation at 37°C. Among the miswaks used, root of the peelu tree in both packing and without packing exhibited strong antimicrobial effect against all three tested microorganisms. However miswak taken from the stem of the peelu and neem tree did not show any antimicrobial activity against all three types of the tested microorganisms. Miswak taken from the root of the peelu tree exhibited antimicrobial activity against all the common oral pathogens and could be a good oral hygiene tool in combating dental caries

Seasonal variation and recent status of typhoid fever in a tertiary care hospital

In many developing countries, typhoid fever is endemic. Early diagnosis is important for appropriate treatment on time. Typhi dot is important diagnostic tool for early diagnosis of typhoid fever. In the current study from February, 2012 to May 2014, samples from 19433 febrile cases referred to Dow Diagnostic Research and Reference lab were screened for serum IgM and IgG for Salmonella typhi by Typhi dot test. Results showed that about 12501 patients had both IgM and IgG antibodies against Salmonella typhi. Male patients were more commonly affected than female patients. Highest frequency of positive cases was seen during the month of October. Age group >15-44 was most likely affected among different age groups. This study suggested that still we are far behind in improving sanitation and elimination of such controllable disease. Therefore prompt measures should be taken to control the disease especially during hot season by vaccination, improved sanitation and education as well

Prevalence and clinical profile of insulin resistance in young women of poly cystic ovary syndrome: a study from Pakistan

Objective: The aim was to estimate the prevalence of Insulin Resistance [IR] in Poly Cystic Ovary Syndrome [PCOS] and analyze its clinical parameters

Methodology: This observational study was conducted at Dow University Hospital during June 2011 till May 2012. Patients of PCOS were selected, an anthropometric measurement, examination and fasting blood test for sugar [FBS] and insulin was performed. Data was collected on pre designed questionnaire, was analyzed by SPSS version 16

Results: Forty-six cases of PCOS were included in the study. Prevalence of IR was 34.78%. Mean age of patients was 23.72 +/- 4.37 years. Waist Hip Ratio [WHR] was raised in 42 [91.30%], acanthosis was found in 26[56.50%], impaired FBS was seen in 9 [19.6%] and raised fasting insulin in 16 [34.8%] patients. There was significant association between acanthosis and WHR [0.044] and between acanthosis and FBS [0.008]. Correlation studies between parameters showed a significant correlation between Waist and Hips [0.93], similarly Waist and WHR showed positive correlation [0.59], at p < 0.01. Significant positive correlation was also found between waist and FBS [0.32] and FBS and WHR [0.378]

Conclusion: Acanthosis nigrican, raised WHR and FBS are significant parameters for insulin resistance in cases of Poly Cystic Ovary Syndrome [PCOS]

Current pattern of bloodstream infections in a tertiary care hospital of Karachi and clinical significance of positive blood cultures

Blood stream infections [BSI] [septicemias] require prompt empirical therapy based on awareness of the drug susceptibility profiles of locally prevalent pathogens isolated. Department of Pathology Dow University of Health Sciences, Karachi, in collaboration with Dr Essa's Diagnostic Centre, from July to November 2008. A cross-sectional prospective study. 324 consecutive blood cultures from patients coming to Civil Hospital Karachi were scrutinized for bacterial isolates and their antibiotic sensitivity profiles were done. A total of 100 [30.9%] specimens were found positive, of these 78% isolates were gram negative bacteria and 22% gram positive cocci. Salmonella typhi, seen less often in adults [20.5%], was the predominant pathogen in children [82.5%]. Escherichia coli, Staphylococcus aureus, Pseudomonas and Klebsiella were isolated in neonates, and Klebsiella, Pseudomonas, Enterobacter and Acinetobacter were identified among adult patients. While all S.typhi isolates were sensitive to cefixime, ceftriaxone and the fluoroquinolones and increasingly amenable to chloramphenicol, of significance was the percentage of other multidrug resistant bacterial isolates. Methicillin resistant Staphylococcu aureus was isolated from one case. The results were analysed by applying SPSS version 16 to derive p value. Amikacin, carbapenems, cefoperazone+sulbactam, fosfomycin and pipericillin+tazobactam are currently the only available drugs still active in-vitro on blood isolates, judicious use of antibiotics focused on the compliance and formation of antibiotic policy guide lines is highly recommended

experimental status of fine needle aspiration cytology of head and neck lesions in a tertiary care scenario

Fine Needle Aspiration Cytology being a minimally invasive technique, is particularly suitable in the sensitive head and neck area where an incisional biopsy may cause problems. FNAC can obviate the need for surgery if the lesion is shown to be non neoplastic or if it confirms suspected metastatic or recurrent tumor. A preoperative cytological diagnosis of a primary neoplasm may allow more rational planning of surgery. This descriptive observational study was carried out at the Department of pathology, DDRRL Dow University of Health Sciences and Sindh Government Qatar hospital during the year 2008-09. A total of 518 consecutive FNAC's of head and neck lesions were included. Out of this 356 cases were from cervical lymphnodes, 124 cases were from thyroid and 38 cases from salivary glands. A total of 45 cases were found malignant and 473 cases were reported as benign lesions

Microcephaly: tracing the evolutionary lineage of ASPM gene

Microcephaly, in the form of congenital autosomal recessive disorder [MCPH], is characterized by the reduced occipital frontal head circumference >3 standard deviation of otherwise normal population of matching age and sex. The disease is primarily associated with mild to severe mental retardation. Earlier studies have unravelled that among Pakistani population, mutations in ASPM gene is strongly associated in MCPH. In the present study, we have explored the ancestral root of this disease and the process involved in its evolution using tools of bioinformatics. Experimental Methods: cDNA gene and protein sequences of ASPM gene were retrieved from NCBI database and subjected to the non-redundant BLAST. Consensus phylogenetic tree was developed after multiple sequence alignment and bootstrapping of the protein sequences of ASPM gene from different mammals using Neighbour Joining method, selecting non mammals as an out group. Comparisons of the gene synteny and exon and intron patterns of ASPM gene were also undertaken to investigate chromosomal changes during the course of human evolution. Different statistical evolutionary models namely, Codon Based Z test and Maximum Composite Likelihood Estimate were used in order to estimate the nature of nucleotide substitution and the type of selection pressure the gene has undergone. Phylogenetic tree based on ASPM gene clearly segregated all non mammalian members as an out group. Mammalian in group holds the established evolutionary lineage, based on morpho-genetic attributes of mammalian evolution, segregating monotremes at the beginning followed by the members of rodentia and finally radiation of the primates including humans. Orientation of the ASPM gene remains conserved between human and chimpanzee, however, it was found reversed along with two flanking genes, a zinc finger binding domain 41 and coagulation factor XIII, which suggest relatively recent event of gene inversion. Some earlier and, in comparison, more intricate chromosomal changes have also been detected among the lower order of mammals. Aligning ASPM gene exons with the primates and lower order mammals indicates transitional bias of mutation over transversion [R value= 1.563]. Holistically, codon based Z test revealed positive selection pressure on of ASPM gene from rodentia to primates. Briefly, the studies highlights the evolutionary events of ASPM gene in mammals especially primates including humans. Further studies in connection to correlating the cranial cavity size and ancestral gene sequences and in depth sequence comparison would be more insightful in this regard and studies in this connection are ongoing and will be reported shortly